Hii,
I am currently using RIPA buffer for the extraction of proteins from neuronal cells. But, getting very faint bands on the blot. Initially, I was doubtful about reagents and the protocol which I am following in western blot. Recently, I did western with HEK cells and glioblastoma cells. Interestingly I got very good bands on the blot. So reagents and methods are not at all a problem. Now, I have doubts about the extraction buffer. I have read that the RIPA buffer contains SDS, which is a harsh detergent. It degrades proteins. So If I use any other Lysis/IP buffer without SDS, will it improve my western results?
https://www.researchgate.net/topic/Lysis-Buffer
There are many RG discussions about the lysis buffer (above link) that may be helpful for your purpose. Alternatively, you could explore method papers using a literature search.
- Badges
- Science method